基因重组靶向结合纤维蛋白的脑源性神经营养因子的制备与生物学活性检测

The Preparation and Bioactivity Assessment of Gene Recombinant Fibrin-binding Brain Derived Neurotrophic Factor

本研究用大肠杆菌表达蛋白质技术制备基因重组脑源性神经营养因子(FBD-BDNF),对该FBD-BDNF的生物学活性和特异结合纤维蛋白的能力进行检验。我们使用大肠杆菌表达目的蛋白,对大肠杆菌表达出的目的蛋白包涵体进行纯化、复性,并通过PAGE胶观察纯度;用ELISA法观察FBD-BDNF与纤维蛋白的结合能力,测定其促进DRG细胞神经轴突生长的能力。本研究成功制备出较纯的FBD-BDNF,结果表明,它能够特异结合纤维蛋白,具有与BDNF相似的神经营养功能,在神经组织工程中具有良好的应用前景。

Using the E. coli, we fabricated the gene reconstructed brain derived neurotrophic factor with a fibrin bind- ing domain （FBD-BDNF）. We then tested the neurotrophic bioactivity and fibrin-binding ability of the FBD-BDNF. The E. coli was used to express the recombinant protein. The inclusion body was purified with column chromatogra- phy and renaturated to construct the right 3D formation. In this study, we successfully fabricated the FBD-BDNF and tested the binding ability and neurotrophic activity. The results demonstrated that FBI^BDNF had special bind- ing ability of fibrin and significant neurotrophic activity for DRG cells. FBD-BDNF could have a promising application prospect in nerve tissue engineering.